Definitive Hematopoiesis in Zebrafish

描述（由申请人提供）：造血干细胞（HSC）生物学影响公共卫生，因为血液疾病患者接受照射以消耗恶性细胞，然后移植健康的HSC以重建其血液。虽然我们对哺乳动物造血干细胞的细胞和功能特性了解很多，但对调控其诱导的遗传途径了解相对较少。斑马鱼作为一个模型系统的发展，允许通过公正的前向遗传筛选方法在脊椎动物中发现基因的无与伦比的能力。最近的转基因技术结合外部受精和斑马鱼胚胎的透明性，可以观察到HSC规范和干细胞生态位形成的事件。这将解决许多长期存在的关于调节干细胞个体发生的分子途径的问题。我们建议进一步表征Notch信号突变体、mindbomb、先锋移植技术在鱼类中的造血表型，并在Notch通路中构建导致HSC规范的分层网络。为了发现HSC诱导所需的新基因，我们将使用已经发现7个新突变体的诱变策略。本K01应用的总体目标是利用斑马鱼的优势，从遗传学上解剖Notch通路在HSC诱导中的作用，并发现控制HSC规范的新基因。这些研究可能阐明新的临床相关基因从鱼转化为人类患者。这项研究将在儿童医院的Leonard Zon医学博士的实验室进行，候选人Caroline Erter Burns博士是该医院的研究员。Zon博士是造血、干细胞和斑马鱼领域的杰出领导者，并将为上述目标提供指导和资源。候选人有医学系的机构承诺。K01奖将帮助伯恩斯博士实现她的目标，成为学术环境中研究干细胞个体发生遗传学的独立研究员。

DESCRIPTION (provided by applicant): Hematopoietic stem cell (HSC) biology impacts public health in that patients with blood disorders undergo irradiation to deplete malignant cells followed by transplantation of healthy HSCs to reconstitute their blood. Although much is known about HSC cellular and functional properties in mammals, relatively little is understood about the genetic pathways regulating their induction. The development of zebrafish as a model system has allowed for the unrivaled ability to discover genes in vertebrates through unbiased forward genetic screening approaches. Recent transgenic technology combined with the external fertilization and transparency of zebrafish embryos allows events in HSC specification and stem cell niche formation to be observed. This will address many long-standing questions regarding the molecular pathways regulating stem cell ontogeny. We propose to further characterize the hematopoietic phenotype in the Notch signaling mutant, mindbomb, pioneer transplantation technology in fish, and build a hierarchical network in the Notch pathway leading to HSC specification. To discover new genes required for HSC induction, we will use a mutagenesis strategy that has already uncovered 7 new mutants. The overall goal of this K01 application is to use the advantages of zebrafish to genetically dissect the role of the Notch pathway in HSC induction and to discover new genes governing HSC specification. These studies may elucidate new clinically relevant genes that translate from the fish into human patients. The research will be conducted in the laboratory of Leonard Zon, M.D. at Children's Hospital where the candidate, Caroline Erter Burns, Ph.D. is a fellow. Dr. Zon is a preeminent leader in the hematopoiesis, stem cell, and zebrafish fields and will provide mentorship and resources for the aims described. The candidate has institutional commitment from the Department of Medicine. A K01 award will aid Dr. Burns in achieving her goal of becoming an independent researcher in an academic setting studying the genetics of stem cell ontogeny.